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The ultimate objective of this project is to discover new and structurally diverse hybrid isoprenoid (HI) antibiotics from marine actinomycetes. The research focuses on an unusual group of marine actinomycetes that were discovered in the PIs laboratory and have tentatively been called MAR4. For reasons that are not yet entirely clear, MAR4 strains have proven to be a rich source of HI secondary metabolites, which are generally rare among bacteria. This proposal takes a unique approach that couples creative culturing techniques and poorly studied environmental resources with molecular approaches to rapidly identify strains capable of producing HI secondary metabolites. These strains then become the focus of detailed structure elucidation and biosynthetic studies that will reveal the mechanisms by which the compounds are assembled. The enzymes discovered as part of these studies subsequently have the potential to be used as biochemical tools to create additional structural diversity. The proposed studies target the discovery of a wide range of HI secondary metabolites including compounds with N and C-prenylation, prenylated pyroles, phenazines, and peptides. The results may include the discovery of diverse new chemical structures within the broad HI class. Major objectives include detailed phylogenetic analyses of MAR4 strains, which will expand our understanding of the diversity of bacteria that produce HIs. A related objective is the biochemical characterization of gene clusters associated with HI biosynthesis and the cloning of specificenzymes for use as biochemical tools. The project goals remained the same during this reporting period. These include an assessment of the diversity of MAR4 marine actinomyetes in ocean sediments collected off the coast of California using both culture and culture independent methods. Goals include the collection of new sediment samples and attempts to draw correlations between the abundance of MAR4 strains and environmental parameters. Additional goals include the identification of hybrid isoprenoid (HI) secondary metabolites produced by MAR4 strains and analyses of the relationships between MAR4 phylotype and the suite of secondary metabolites produced. We continue to work on pathways associated with HI biosynthesis, with a focus on strain CNH-189, which produces the unusual merochlorin class of compounds. |
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RXMCDM: 金币+1, 多谢应助 2014-10-12 12:00:25
bxzc123: 金币+35, 翻译EPI+1, ★★★很有帮助, 谢谢 2014-10-12 19:30:02
bxzc123(RXMCDM代发): 金币+15 2014-10-13 20:14:47
RXMCDM: 金币+1, 多谢应助 2014-10-12 12:00:25
bxzc123: 金币+35, 翻译EPI+1, ★★★很有帮助, 谢谢 2014-10-12 19:30:02
bxzc123(RXMCDM代发): 金币+15 2014-10-13 20:14:47
| 这个项目的最终目标是从海洋放线菌中发现新的和结构多样化的混合类异戊二烯。这项研究的对象是一种特殊的海洋放线菌，它是由PIs实验室发现的，暂定取名为MAR4，主要原因是目前还没有完全弄清楚，MAR4系被证明含有丰富的HI次生代谢产物，这在一般的细菌中时很罕见的，这个方案的特别之处在于，兼具创造行的培养技术，以及在恶劣的研究环境下能够利用分子分析方法迅速的分析出其制造混合类异戊二烯二级产物的能力，这样就能对MAR4系进行具体结构分析和生物合成研究，进而通过这些结果对其进行合成。随后，在这个研究中发现的酶很有作为生化工具创造其它结构复杂物质的潜力。这个项目的研究目的是发现一种包含N和c-异戊烯、异戊吡咯、吩嗪 、肽的泛HI二级产物，结果也许会包含HI家族中的结构多样化的新物质。主要目标还包括对MAR4系的具体进化分析，这能够帮助我们了解更多能够产生HIs的多样化细菌，相关结果是与HIs合成相关基因的生化表征和以及克隆出这种能够作为生化工具的酶，这个项目还在研究阶段的是，利用对从加利福尼亚海岸沉积物中收集的MAR4放线菌进行培养或者单独培养的方式对其多样性进行评估，目标包括收集新的泥沙样本，以试图找出MAR4系丰富程度与环境参数的关系。另外还包括鉴定MAR4系所产生的HI次生代谢物和分析MAR4放线菌和生产次生代谢产物的工具的关系。我们将继续研究HIs合成的相关方法，主要集中在对CNH-189系的研究，它能够产生一种特殊的酶。 |
2楼2014-10-12 10:46:29
